Development of analytical methods for glycosaminoglycans from biological sources

生物来源糖胺聚糖分析方法的开发

基本信息

批准号：
10545230
负责人：
Zhangjie Wang
金额：
$ 98.88万
依托单位：
GLYCAN THERAPEUTICS CORPORATION
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-15 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10545230
关键词：
Animals Biological Biological Process Biomedical Research Blood coagulation Brain Businesses Cell Differentiation process Chondroitin Chondroitin Sulfates Clinical Trials Development Disaccharides Embryonic Development Enzymes Glycosaminoglycans Goals Growth Heparin Heparin Lyase Heparitin Sulfate Human Income Inflammatory Response Injury Label Letters Low-Molecular-Weight Heparin Measurable Messenger RNA Methods Modification Mus Neoplasm Metastasis Neurodegenerative Disorders Neurons North Carolina Oligosaccharides Parasitic infection Patients Phase Plasma Polysaccharides Reaction Reporting Research Resolution Role Sales Sampling Series Services Small Business Innovation Research Grant Solid Source Structure Sulfate Therapeutic Time Tissues Tracer Universities Validation Viral Work analytical method base brain tissue cost estimate depolymerization design diagnostic tool disease diagnosis dosage experience experimental study improved in vivo innovation mouse model phase 1 study phase 2 study quality assurance quinoline subcutaneous success tool trend

项目摘要

This SBIR phase II proposal is aimed to develop a LC-MS/MS based method to analyze heparan sulfate and chondroitin sulfate with sub-nanogram quantities. Heparan sulfate and chondroitin sulfate are highly sulfated polysaccharides displaying a wide range of biological functions. There is a strong demand for a quantitative and sensitive method to analyze heparan sulfate and chondroitin sulfate. In this proposal, we propose to achieve the quantitation of heparan sulfate and chondroitin sulfate using 13C-labeled calibrants. To this end, heparan sulfate and chondroitin sulfate are depolymerized into disaccharides by heparin lyases and chondroitin ABCase. The composition analysis of the resultant disaccharides is accomplished by a LC-MS/MS method. In addition, we plan to use 13C-labeled heparan sulfate octasaccharide calibrants to determine the levels of 3-O-sulfated heparan sulfate. The crucial innovation in our proposed method is to add “absolute quantitation” capability using 13C-labeled disaccharide calibrants and 13C-labeled octasaccharide calibrants. Aim 1 is designed to demonstrate the ability to quantitative the level of 3-O-sulfated heparan sulfate. We will focus on analyzing the analysis of 3-O-suflated heparan sulfate from plasma samples and from human brain tissues with neurodegenerative diseases. Aim 2 is aimed to develop a method for quantitative disaccharide analysis of chondroitin sulfate. We will synthesize six 13C- labeled chondroitin sulfate disaccharide calibrants to carry out the studies. We will also conduct the analysis for chondroitin sulfate from human brain tissues from neurodegenerative diseases. The completed heparan sulfate and chondroitin sulfate analysis from plasma and solid tissues will demonstrate the robustness of the method. The success of this project will provide a method to analyze heparan sulfate and chondroitin sulfate with sensitivity and accuracy for biologists.

该 SBIR II 期提案旨在开发一种基于 LC-MS/MS 的方法来分析硫酸乙酰肝素和硫酸软骨素的量为亚纳克级。和硫酸软骨素是高度硫酸化的多糖，具有广泛的迫切需要一种定量且灵敏的方法。为了分析硫酸乙酰肝素和硫酸软骨素，在本提案中，我们建议：使用 13C 标记实现硫酸乙酰肝素和硫酸软骨素的定量为此，硫酸乙酰肝素和硫酸软骨素被解聚成。肝素裂解酶和软骨素 ABCase 的二糖成分分析。所得二糖是通过 LC-MS/MS 方法完成的。计划使用 13C 标记的硫酸乙酰肝素八糖校准物来测定我们提出的方法的关键创新是3-O-硫酸化硫酸乙酰肝素的水平。使用 13C 标记的二糖校准物添加“绝对定量”功能，以及 13C 标记的八糖校准物旨在证明其能力。我们将重点分析 3-O-硫酸化硫酸乙酰肝素的水平。分析血浆样品和人脑中的 3-O-硫酸乙酰肝素目标 2 旨在开发一种方法来治疗患有神经退行性疾病的组织。硫酸软骨素的定量二糖分析我们将合成六种13C-。我们还将标记硫酸软骨素二糖校准物来进行研究。对人脑组织中的硫酸软骨素进行分析完整的硫酸乙酰肝素和硫酸软骨素。血浆和固体组织的分析将证明该方法的稳健性。该项目的成功将为分析硫酸乙酰肝素和硫酸软骨素对生物学家来说具有敏感性和准确性。